Ribosomal display is a cell-free system using the principle of coupling phenotype (protein) to genotype (gene) for the in vitro selection of proteins from large libraries (Mattheakis et al., Proc. Natl. Acad. Sci. USA, 91:9022-9026 (1994); Hanes et al., Proc. Natl. Acad. Sci. USA, 94:4937-4942 (1997); He et al., Nucleic Acids Res. 25(24):5132-4 (1997)). Prokaryotic and eukaryotic ribosomal display systems have been used for the selection of peptides, single-chain antibodies (e.g., scFvs), enzymes, stable protein scaffolds and other ligand-binding domains (He et al., Brief Funct. Genomic Proteomic, 1(2):204-12 (2002)). The methods described herein increase the efficiency of ribosomal display methods for producing and selecting proteins of interest from very large libraries (e.g., between about 103 to 105 members, or about 105 to 1016 members).